interleukin-I Beta Research Articles

Alternative skin sensitization prediction and risk assessment using proinflammatory biomarkers, interleukin-1 beta (IL-1β) and inducible nitric oxide synthase (iNOS)

ABSTRACTAs an alternative to animal tests for skin sensitization potency and risk assessment, cell viability and biomarkers related to skin sensitization were analyzed in THP-1 human monocytic leukemia cells. Cell viabilities of 90% (CV90) and 75% (CV75) were determined for 24 selected test chemicals. Further biomarkers related to skin sensitization were also determined under equivalent comparative conditions. In cell viability analyses, potent skin sensitizers exhibited high cytotoxicity, but non-sensitizers did not display this tendency. In biomarker analyses, interleukin-I beta (IL-1β), inducible nitric oxide synthase (iNOS), IL-1β+iNOS, and THP-1 IL-1β+Raw 264.7 IL-1β were found to be suitable for prediction of skin sensitization potency following classification as either skin sensitizers or non-sensitizers (accuracies of 91.7%, 87.5%, 83.3%, and 82.6%, respectively). A significant positive correlation was found between biomarkers and skin sensitization potency, with a correlation coefficient (R) of 0.7 or more (correlation coefficients of 0.77, 0.72, 0.7, and 0.84, respectively). Finally, the skin sensitization potency effective threefold concentration (EC) 3% was predicted using a biomarker equation, with resulting prediction rates (match rate with actual data) of 58.3%, 54.2%, 62.5%, and 60.9%, respectively. The prediction accuracy for the EC3 value obtained from animal data was calculated as 83.3%, 79.2%, 79.2%, and 73.9%, respectively. Thus, these biomarkers, IL-1β and iNOS, may be alternatively used to predict skin sensitization potency and risk assessment.

Comparative Aspects of the Endotoxin‐ and Cytokine‐Induced Endocrine Cascade Influencing Neuroendocrine Control of Growth and Reproduction in Farm Animals

Disease in animals is a well-known inhibitor of growth and reproduction. Earlier studies were initiated to determine the effects of endotoxin on pituitary hormone secretion. These studies found that in sheep, growth hormone (GH) concentration was elevated, whereas insulin-like growth factor-I (IGF-I) was inhibited, as was luteinizing hormone (LH). Examination of the site of action of endotoxin in sheep determined that somatotropes expressed the endotoxin receptor (CD14) and that both endotoxin and interleukin-I beta activated GH secretion directly from the pituitary. In the face of elevated GH, there is a reduction of IGF-I in all species examined. As GH cannot activate IGF-I release during disease, there appears to be a downregulation of GH signalling at the liver, perhaps related to altered nitration of Janus kinase (JAK). In contrast to GH downregulation, LH release is inhibited at the level of the hypothalamus. New insights have been gained in determining the mechanisms by which disease perturbs growth and reproduction, particularly with regard to nitration of critical control pathways, with this perhaps serving as a novel mechanism central to lipopolysaccharide suppression of all signalling pathways. This pathway-based analysis is critical to the developing novel strategies to reverse the detrimental effect of disease on animal production.

Genistein reduces the production of proinflammatory molecules in human chondrocytes

Previously, we reported that cartilage is an estrogen receptor (ER) positive tissue and that mRNA levels of ERβ increase in postmenopausal women with osteoarthritis. Based on our findings and those of other investigators, we hypothesized that local rather than circulating estrogen levels negatively affect chondrocyte metabolism and that selective ER modulators (SERM) augment cartilage health. To test the latter part of our hypothesis, we explored the role of genistein, a naturally occurring SERM with high affinity to bind ERβ, in inhibiting the lipopolysaccharide (LPS)-stimulated cyclooxygenase (COX)-2 in chondrocytes. Primary cultures of normal human chondrocytes were treated with three levels of genistein (0, 50, and 100 μM). After 1 h, the genistein-treated cells were stimulated by 1 μg/ml LPS for 24 h. Cells were then harvested, and the cytosolic fraction was isolated for assessment of COX-1 and COX-2 protein levels using Western analysis. Nitric oxide (NO), interleukin-I beta (IL-1β), and human cartilage glycoprotein 39 (YKL-40) production was also measured in cell supernatants. NO and IL-1β were measured as markers of inflammation, and YKL-40 was assessed as a marker of cartilage catabolism. Genistein had no significant effect on either YKL-40 or IL-1β levels. Our data indicate that the LPS-stimulated increases in COX-2 protein level and NO in supernatant are reduced by pretreatment of genistein, whereas COX-1 protein level is not affected by genistein. The ability of genistein to suppress COX-2 but not COX-1 is advantageous because suppressing COX-2 can lead to suppression of proinflammatory molecules. Although genistein suppresses COX-2 production, it does not affect the production of COX-1 enzyme, which is responsible for releasing prostaglandins involved in cellular house-keeping functions such as the maintenance of gastrointestinal integrity and vascular homeostasis.

Interleukin-10 gene -627 allele variants, not interleukin-I beta gene and receptor antagonist gene polymorphisms, are associated with atopic bronchial asthma.

Asthma is an airway hyperresponsive disease characterized by the expression of multiple inflammatory genes, including cytokines. Interleukin-I and interleukin-10 (IL-1 and IL-10) are cytokines that might play a role in the process of inflammation and are therefore considered to be involved in the pathogenesis of bronchial asthma. The aim of this study was to test whether the polymorphisms of the promoter region and exon 5 of the IL-1 gene, intron 2 of the IL-1Ra gene, and -627 nucleotide (C/A) of the IL-10 gene could be genetic markers for the susceptibility of bronchial asthma. A normal control group made up of 47 healthy volunteers and 117 patients with bronchial asthma were examined in this study. We analyzed the variable number of tandem repeats at intron 2 of the IL-1Ra gene for the polymorphisms by polymerase chain reaction (PCR). PCR-based restriction analysis of the IL-1 gene polymorphisms of the promoter region and exon 5 was carried out by the endonucleases Ava I and Taq I, respectively. The IL-10 gene -627 C/A polymorphisms were investigated by PCR-based restriction analysis. The distribution of CC homozygotes in the IL-10 gene was significantly lower in asthma patients than in controls (P=0.013, OR=3.599, 95% CI=1.240 approximately 10.441). The polymorphisms studied in the IL-1 genes did not reveal any significant association with bronchial asthma when compared with the control group (promoter region by chi-square test, P=0.627; exon 5 region by Fisher's exact test, P=0.403). Only two alleles of the IL-1Ra gene corresponding to one and two copies of an 86-base pair sequence repeat were identified by PCR in the control group. There were three alleles found in the asthmatic patient group. The results revealed no significant differences between normal individuals and asthma patients (P=0.454, Fisher's exact test). The IL-10 gene -627 "A" allele is an associated risk factor of developing atopic asthma.

Pathogenesis of joint damage in rheumatoid arthritis: evidence of a dominant role for interleukin-I.

Chronic arthritis is characterised by persistent joint inflammation and concomitant joint destruction. Although joint swelling is a major clinical feature, destruction of bone and cartilage may be dissociated from inflammation. It is therefore important to understand fully all elements of the destructive process. Tumour necrosis factor (TNF) and interleukin-I (IL-I) are considered pivotal cytokines in the process of human rheumatoid arthritis (RA), with a claimed cascade of TNF inducing most of the IL-I production. Studies in experimental models have revealed that TNF is indeed a pivotal cytokine in acute joint swelling, yet IL-I beta is the dominant cartilage destructive cytokine and its production may occur independently of TNF alpha. This was found with anti-TNF/IL-I neutralising antibodies and the observations were recently supported by similar findings in arthritis models in TNF and IL-I knock-out mice. In RA, early clinical studies suggested a correlation between levels of IL-I beta and measures of joint damage. In vitro studies have also demonstrated regulatory effects of IL-I beta on both cartilage degradation and cartilage invasion by synoviocytes. A randomised clinical trial has suggested a significant reduction in the rate of joint damage following IL-I beta inhibition by IL-I receptor antagonist. Clinical trials of TNF alpha blockade have demonstrated a marked reduction in the clinical manifestations of inflammation but, to date, an effect on the rate of joint damage awaits confirmation.

Increased release of tumor necrosis factor-alpha and interleukin-6 in women with the syndrome of hemolysis, elevated liver enzymes, and low platelet count.

Complement is activated in preeclampsia and complement products are known to activate macrophages. The aim of this study was to determine whether the macrophage derived cytokines, interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha, are released in patients with a form of severe preeclampsia characterized by the syndrome of hemolysis, elevated liver enzymes, and low platelet count (HELLP syndrome). Complement activation and plasma levels of cytokines were studied in 11 women with HELLP syndrome and in 11 controls with uncomplicated pregnancies. To further evaluate the connection between complement activation and cytokine release an in vitro study on heparinized whole blood incubated with recombinant C5a was performed. In the HELLP group, complement anaphylatoxin C5a was increased in plasma at delivery (p < 0.01) and one day after delivery (p < 0.05), terminal C5b-9 complement complex was elevated in plasma at delivery (p < 0.001) and one day after (p < 0.01), plasma levels of interleukin-6 were increased one day after delivery (p < 0.01), and plasma concentrations of tumor necrosis factor-alpha were elevated at delivery (p < 0.01), compared with corresponding levels in controls. All parameters normalized within one week. Interleukin-I beta did not differ between the groups. In vitro, recombinant C5a incubated in whole blood gave a dose-dependent release of interleukin-6. No increased release of interleukin-1 or tumor necrosis factor-alpha was seen after incubation. Since cytokine release occurs in severe preeclampsia, inflammatory mechanisms may participate in the pathophysiology of severe preeclampsia.

Adhesion of Sickle Red Blood Cells and Damage to Interleukin-1β Stimulated Endothelial Cells Under Flow In Vitro

Two factors that are hypothesized to contribute to vasoocclusive crises in sickle cell anemia are increased sickle red blood cell-endothelial cell interactions and damage to endothelium. Despite considerable study, the mechanisms by which erythrocyte-endothelial interactions occur and the role of endothelial damage have not yet been fully elucidated. In this report, we demonstrate that adhesion and damage may be related in a model of vasoocclusion in sickle cell anemia. Phase contrast microscopy coupled to digital image processing was used to determine the adhesion of sickle red blood cells to 1-, 4-, and 24-hour interleukin-I beta (IL-1 beta) stimulated endothelial calls in a parallel plate flow chamber. Morphological alterations to activated endothelial cells after the perfusion of sickle erythrocytes were also identified. Pretreatment of monolayers with 50 pg/mL of IL-1 beta for 1, 4, and 24 hours caused approximately 16-fold increases in adhesion of sickle cells to activated endothelium at all time points. Results with an Arginine-glycine aspartic acid (RGD) peptide and monoclonal antibodies indicated a role for three different endothelial cell receptors: alpha v beta 3 after 1 hour of IL-1 beta stimulation; E-selectin after 4 hours of IL-1 beta stimulation; and vascular cell adhesion molecule-1 after prolonged exposure to cytokines. Perfusion of sickle, but not normal, erythrocytes resulted in alteration of endothelial morphology. Approximately 6% to 8% damage was observed on 4- and 24-hour IL-1 beta stimulated endothelial cells after the perfusion of sickle cells. Damage to 24-hour activated endothelial cells showed a positive correlation (r = .899) with the number of adherent sickle erythrocytes.

Copper addition prevents the inhibitory effects of interleukin 1-beta on rat pancreatic islets.

Since copper [Cu(II)] is a necessary cofactor for both intra-mitochondrial enzymes involved in energy production and hydroxyl scavenger enzymes, two hypothesised mechanisms for action of interleukin-I beta (IL-1 beta), we studied whether Cu(II) addition could prevent the inhibitory effect of IL-1 beta on insulin release and glucose oxidation in rat pancreatic islets. Islets were incubated with or without 50 U/ml IL-1 beta, in the presence or absence of various concentrations of Cu(II)-GHL (Cu(II) complexed with glycyl-L-histidyl-L-lysine, a tripeptide known to enhance copper uptake into cultured cells). CuSO4 (1-1000 ng/ml) was used as a control for Cu(II) effect when present as an inorganic salt. At the end of the incubation period, insulin secretion was evaluated in the presence of either 2.8 mmol/l (basal insulin secretion) or 16.7 mmol/l glucose (glucose-induced release). In control islets basal insulin secretion was 92.0 +/- 11.4 pg.islet-1 h-1 (mean +/- SEM, n = 7) and glucose-induced release was 2824.0 +/- 249.0 pg.islet-1 h-1. In islets pre-exposed to 50 U/ml IL-1 beta, basal insulin release was not significantly affected but glucose-induced insulin release was greatly reduced (841.2 +/- 76.9, n = 7, p < 0.005). In islets incubated with IL-1 beta and Cu-GHL (0.4 mumol/l, maximal effect) basal secretion was 119.0 +/- 13.1 pg.islet-1 h-1 and glucose-induced release was 2797.2 +/- 242.2, (n = 7, p < 0.01 in respect to islets exposed to IL-1 beta alone).(ABSTRACT TRUNCATED AT 250 WORDS)

Possible regulation of soluble ICAM-1 levels by interleukin-1 in a sub-set of breast cysts.

There are 2 main groups of breast cysts characterized by their intracystic sodium-to-potassium ratios. Women who have intracystic Na/K < 3 may have a higher risk of developing breast cancer than those who have intracystic Na/K > 3. The cell adhesion molecule, ICAM-I, has been shown to be inducible by interleukin-I (IL-I). Furthermore, ICAM-I may be involved in the progression of metastasis in certain malignancies. The aim of this study was to measure intracystic soluble ICAM-I (sICAM-I) and IL-I alpha/beta levels and to assess the relationships between these substances. Wide-ranging concentrations of sICAM-I, IL-I alpha and IL-I beta were found in breast-cyst fluid. The high-electrolyte-ratio group had intracystic levels of sICAM-I and IL-I beta which were significantly higher than those of the low-electrolyte-ratio group. No significant difference was found between IL-I alpha concentrations in the 2 groups of breast cysts. Strong positive correlations were found between sICAM-I and IL-I alpha, sICAM-I and IL-I beta, and IL-I alpha and IL-I beta. The significance of the higher concentrations of sICAM-I in the high-electrolyte-ratio group remains to be determined, but the results of this study suggest that sICAM-I levels in the high-electrolyte-ratio group may be regulated in part by IL-I alpha and IL-I beta.

Serum Interleukin-1 Beta is Increased in Cluster Headache

We measured serum interleukin-1 beta (IL-1 beta) in 24 episodic cluster headache (CH) patients and 45 normal controls using a specific ELISA method. There was an increase in IL-1 beta in all CH patients compared to controls. IL-1 beta was further increased during the ictal phase of CH compared to patients between attacks and normal individuals. Between attacks, IL-1 beta was also significantly increased compared to controls. We suggest that these results represent an activation of the immune system in CH.